LOCUS       sequence-13330-e        4147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optogenetic tool for light modulation of cellular cAMP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4147)
  AUTHORS   Stierl M, Stumpf P, Udwari D, Gueta R, Hagedorn R, Losi A, Gartner 
            W, Petereit L, Efetova M, Schwarzel M, Oertner TG, Nagel G, Hegemann
            P
  TITLE     Light modulation of cellular cAMP by a small bacterial 
            photoactivated adenylyl cyclase, bPAC, of the soil bacterium 
            Beggiatoa.
  JOURNAL   J Biol Chem. 2011 Jan 14. 286(2):1181-8.
  PUBMED    21030594
REFERENCE   2  (bases 1 to 4147)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2011 Jan 14. 286(2):1181-8."
COMMENT     Sequence Label: pGEM-HE-h_bPAC_cmyc
FEATURES             Location/Qualifiers
     source          1..4147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..53
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     45..62
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     46..62
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     69..88
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        69..87
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           119..161
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1258..1287
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     3'UTR           1311..1436
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1557..1575)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1558..1575)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1593..1609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1593..1609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1606..1628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1617..1633
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1641..1647,1648..1665,1666..1671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1686..1707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1824..1841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1995..2583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2075..2094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2754..3545,3546..3614))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3377..3396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3615..3719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3787..3805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3843..3865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3965..3984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
       61 gtgaattgta atacgactca ctatagggcg aattaattcg agctcggtac ccagcttgct
      121 tgttcttttt gcagaagctc agaataaacg ctcaactttg gcagatcaat tccccgggga
      181 tccaagcttg ccaccatgat gaagcggctg gtgtacatca gcaagatcag cggccacctg
      241 agcctggaag agatccagcg gatcggcaag gtgtccatca agaacaacca gcgggacaac
      301 atcaccggcg tgctgctgta cctgcagggc ctgttcttcc agatcctgga aggcgagaac
      361 gagaaggtgg acaagctgta caagaaaatc ctggtggacg accggcacac caacatcctg
      421 tgcctgaaaa ccgagtacga catcaccgac cggatgttcc ccaactgggc catgaaaacc
      481 atcaacctga acgagaacag cgagctgatg atccagccca tcaagagcct gctgcagacc
      541 atcacccaga gccaccgggt gctggaaaag tacatgcccg ccagagtgat ctacctgatc
      601 aaccagggca tcaaccccct gaccgtggag ccccagctgg tcgagaagat catcttcttc
      661 agcgacatcc tggccttcag caccctgacc gagaagctgc ccgtgaacga ggtggtcatc
      721 ctggtcaacc ggtacttcag catctgcacc cggatcatca gcgcctacgg cggcgaagtg
      781 accaagttca tcggcgactg cgtgatggcc agcttcacca aagagcaggg cgacgccgcc
      841 atccggacca gcctggacat catctccgag ctgaagcagc tgcggcacca cgtggaggcc
      901 accaaccccc tgcacctgct gtacaccggc atcggcctga gctacggcca cgtgatcgag
      961 ggcaacatgg gcagcagcct gaagatggac cacaccctgc tgggggacgc cgtgaacgtg
     1021 gccgccaggc tggaagccct gacaagacag ctgccctacg ccctggcctt tacagccggc
     1081 gtgaagaagt gctgccaggc ccagtggacc ttcatcaacc tgggagccca ccaggtcaag
     1141 ggcaagcagg aagccatcga ggtctacacc gtgaatgagg cccagaagta ctacgacacc
     1201 ctgcagatca cccagctgat ccggcagacc ctggaaaacg acaagccgcg gacgtacgag
     1261 caaaagctga tctcggagga ggacctgtaa gcttgatctg gttaccacta aaccagcctc
     1321 aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt acaaaatgtt
     1381 gtcccccaaa atgtagccat tcgtatctgc tcctaataaa aagaaagttt cttcacattc
     1441 taaaaaaaaa aaaaaaaaaa aaaacccccc cccccccccc cccccccccc ccccctgcag
     1501 gcatgcgttt aaacatttaa attacgtatt aattaagata tcgctagctt gagtattcta
     1561 tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1621 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1681 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1741 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1801 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1861 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1921 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2821 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2881 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2941 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3001 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3061 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3721 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3781 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3841 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3901 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3961 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4021 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     4081 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4141 gctgcaa
//
